Cholesterol as a natural probe for free radical-mediated lipid peroxidation in biological membranes and lipoproteins

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Albert W Girotti, Witold Korytowski

Abstract

We describe a relatively convenient and reliable procedure for assessing the magnitude of free radical-mediated (chain) lipid peroxidation in biological systems. The approach is based on use of radiolabeled cholesterol ([(14)C]Ch) as a probe and determination of well-resolved oxidation intermediates/products ([(14)C]ChOX species), using high performance thin layer chromatography with phorphorimaging detection (HPTLC-PI). In a lipid hydroperoxide-primed liposomal test system treated with ascorbate and a lipophilic iron chelate, the following well-resolved [(14)C]ChOX are detected and quantified: 7α/7β-OOH, 7α/7β-OH, and 5,6-epoxide, their levels increasing with incubation time at 37°C. [(14)C]Ch also serves as an excellent probe for lipid peroxidation in lipoproteins and plasma membranes of mammalian cells. Because this approach utilizes Ch as a natural in situ probe, it eliminates potential artifacts associated with artificial probes such as spin traps and fluorophores.

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Citations

Jul 3, 2018·Photochemistry and Photobiology·Albert W Girotti, Witold Korytowski
Feb 26, 2019·Soft Matter·Benjamin ViadaLidia M Yudi
Mar 9, 2019·International Journal of Molecular Sciences·Shi-Hui LawLiang-Yin Ke
Jul 6, 2017·Biosensors·Jessica Z Kubicek-SutherlandHarshini Mukundan
Apr 24, 2017·Cell Biochemistry and Biophysics·Albert W Girotti, Witold Korytowski
Apr 19, 2020·Cell Biochemistry and Biophysics·Albert W Girotti, Witold Korytowski

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